Phytochemical profiling and anti-inflammatory evaluation of Raphanus raphanistrum subsp. sativus (L.) extracts

Abstract

Dysregulated inflammatory responses may result in various chronic diseases. Traditionally, non-steroidal anti-inflammatory drugs (NSAIDs) have been used for the treatment of inflammatory diseases. However, due to their serious adverse effects when used for long-term, the researcher has developed an interest in natural plant-based remedies. The current study investigated the phytochemical composition and anti-inflammatory activity of Raphanus raphanistrum subsp. sativus (L.), a Brassicaceae member which is traditionally used as a source of nutrition and also against various diseases. The whole plant was collected from Feroza village (District Rahim Yar Khan, Pakistan), shade-dried, powdered, and extracted with two solvents: methanol and dichloromethane (DCM). Preliminary phytochemical screening discovered the presence of alkaloids, cardiac glycosides, terpenoids, and flavonoids, whereas tannins and lipids were absent. Thin-layer chromatography (TLC) and column chromatography analyses yielded multiple fractions rich in bioactive constituents. The DCM extract demonstrated notable in vitro anti-inflammatory activity, inhibiting 53.6% of reactive oxygen species (ROS) generation at 100 µg/mL (IC₅₀ = 93.5 ± 2.4 µg/mL), compared to ibuprofen (73.2 ± 1.4% inhibition, IC₅₀ = 11.2 ± 1.9 µg/mL). In vivo experiments were carried out using carrageenan-induced paw edema in Wistar rats. This further confirmed a significant, dose-dependent inhibition of inflammation, which was comparable to that of diclofenac sodium when used at higher doses. These findings validate the ethnomedicinal use of R. raphanistrum. Our results demonstrated that this plant can be a promising natural source of anti-inflammatory compounds. However, future studies using advanced spectroscopic and mechanistic analyses are required to identify the specific active constituents and explore their pharmacological mechanisms.